Short technical reportDevelopment and characterization of multiplex panels of polymorphic microsatellite loci in giant liver fluke Fascioloides magna (Trematoda: Fasciolidae), using next-generation sequencing approach

- 118 microsatellite loci generated after NGS for Fascioloides magna.
- 36 loci selected after PCR tests and Sanger sequencing.
- 12 polymorphic loci subjected to fragment analysis and for multiplex panels.
- 11 final loci suitable for population genetics of global Fascioloides magna.

The microsatellite markers were designed for the giant liver fluke, Fascioloides magna, veterinary important liver parasite of free-living and domestic ruminants. Due to its geographic distribution (five enzootic regions across USA and Canada, three permanent European foci) and invasive character, F. magna is an interesting model for population genetics. Out of 667 amplicon candidates generated after NGS, 118 provided the best resolution and were tested with PCR analysis. In total, 56 yielded PCR products of expected size and in 36 of them the declared repetitive motif was identified by Sanger sequencing. After fragment analysis, 12 loci were proved to be polymorphic in individuals from one tested European and four North American populations. These loci were selected for setup of multiplex STR assays and utilized in genotyping of larger sample cohort. The outputs of statistical analyses indicate further global application of 11 conclusive loci in population genetics of the parasite.

The eleven microsatellite loci were designed for Fascioloides magna. Their applicability was tested on global populations what ensures the future comprehensive use in population genetics.232